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Please use this identifier to cite or link to this item: http://hdl.handle.net/123456789/9063

Title: Rapid cODlInunication: Linkage mapping of the Mahogany (attractin) locus in cattle and pigs!
Authors: B. Edeal
J. M. Rumph
Issue Date: 2004
Abstract: Polymorphism. Restriction fragment length polymor¬phisms (RFLP) were detected within peR amplification products of the porcine and bovine Mahogany (attractin; ATRN) genes using the restriction enzymes Taq I and MspI, respectively. Primer Description. The common (porcine and bovine) forward and reverse primers were designed based on homologous regions of human and mouse ATRN cDNA sequences (GenBank accession no. ABOl1120 and AF120318, respectively). These primers amplified a re¬gion of the porcine and bovine ATRN gene spanning from exon 5 to the 3' untranslated region. Primer Sequences. Forward primer: 5'-GTGTACAAG-GAGAAGTCAGGAG-3'; reverse primer: 5'-GATC-TATT( Cfr)AAAGTCTAGGCAC-3'. Method of Detection. peR amplification was performed using 50 ng of genomic DNA and 1 U of Taq Gold poly¬merase (Perkin Elmer, Foster City, CA) with the sup¬plied buffer (final concentration of 1.5 mM MgCI2). "Touchdown" thermal cycling used 10 cycles of amplifi¬cation with annealing temperatures beginning at 65°C and decreasing 1 °C per cycle, followed by 30 cycles with an annealing temperature of 55°C. Each cycle had an extension time of 2.5 min. Digestion of the resulting porcine -980-bp product with Taq I revealed a polymor¬phism with two alleles (Figure 1). An 800-bp band is present in Allele A, with band sizes of 500 and 300 bp characterizing Allele B. Additional smaller bands were not visualized on the 4% metaphor (FMC BioProducts, Rockland, ME) gel. Digestion of the resulting bovine -980-bp product with MspI revealed a polymorphism with two alleles (Figure 1) characterized by bands of 550 and 250 bp (A allele) or 550 and 220 and 30 bp (B allele). Additional smaller bands were not visualized on the 4% metaphor gel. Confirmation of the identities of the por¬cine and bovine ATRN peR amplification products was obtained by terminal end sequencing (GenBank acces-sion no. AF194961 and AF194962, respectively). Mendelian Inheritance. Mendelian segregation of por¬cine ATRN was confirmed in eight full-sib families (91 offspring). Mendelian inheritance of bovine ATRN was confirmed in 17 full-sib families (175 offspring). Chromosomal Location. In a sex-averaged analysis us¬ing the three-generation PiGMaP reference families (Ar¬chibald et aI., 1995), ATRN was linked to several micro¬satellite markers on chromosome 17 (SSC17), including 80292, SWl 031, and 80359 with recombination frequen¬cies of 0.09, 0.20, and 0.17 and LOD scores of 5.27,4.17, and 5.58, respectively. In addition, ATRN was linked to a marker within the ENDO locus (.06, 5.81) on SSC17.
URI: http://hdl.handle.net/123456789/9063
Appears in Collections:College of Foods And Agricultural Science

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